جستجوی مقالات مرتبط با کلیدواژه "اسپرماتوگونی" در نشریات گروه "پزشکی"

In vitro proliferation and maintenance of spermatogonial stem cells is one of the treatment options for infertile men. Creating a suitable microenvironment similar to the natural conditions of reproduction of these cells leads to the improvement of culture and spermatogenesis. Therefore, the aim of the present study was to use a 3D silk nanofibrous electrospun scaffold and co-culture with Sertoli cells in order to increase the proliferation of spermatogonial stem cells.

In this experimental study, spermatogonial stem cells were isolated from the testes of 50 2-4-day-old mice (5 samples each time) using two-step mechanical and enzymatic digestion within two weeks. After placing 2×104 cells on the surface of the silk scaffold, they were exposed to culture medium. The viability of these cells was assessed using MTT assay and their binding was evaluated by SEM microscopy. The studied variables, including Stra8, DAZL and Piwill2 specific genes were analyzed by real time PCR and immunocytochemistry.

The viability of cultured cells in the presence of scaffold and Sertoli (91±6), (90±2) was higher than the control group (85±2), (83±8). Real time PCR results confirmed a significant increase in the expression of specific markers of spermatogonial stem cells such as Stra8, Piwill2 and DAZL in cells cultured respectively on scaffold (1.47±6) (1.28±5) (1.43±2) compared to the 2D culture group (1.17±5) (1.05±3) (1.13±7) (p<0.05).

The results showed that silk scaffold in the presence of Sertoli cell co-culture can increase in vitro proliferation of spermatogonial stem cells and can have a positive effect on the viability and proliferation of these cells.

Spermatogonial Stem Cells (SSCs) differentiate into adult germ cells during spermatogenesis and cause male fertility. Maintenance of SSCs and induction of spermiogenesis in vitro may provide a therapeutic strategy to treat male infertility. Moreover, it is possible to use the SSCs for research in the field of biotechnology and regenerative medicine through differentiation of them into other cell lines.

SSCs were isolated enzymatically from digested testicular tissue and then were cultured. The identity of SSCs were confirmed by anti-OCT4, anti-PLZF through immunocytochemistry. SSCs were cultured in differentiated culture medium. Then, expression level of SCP3, Boule, Crem, Protamine 2 genes and also expression level of SCP3, Protamine2 proteins was determined by Real-time PCR and immunocytochemistry, respectively. In addition, the potency of SSCs differentiation into osteoblasts and adipocytes was evaluated by Alizarin Red and Oil Red staining, respectively.

The results showed that SSCs can differentiate into adult germ cells in in vitro condition. Also, under differentiation conditions, these cells can be distinguished from other cell lines.

The results of this study showed that, given the differentiation potency of SSCs, these cells can be used for the treatment of male infertility, as well as cell therapy and regenerative medicine.

Spermatogonial stem cells (SSCs) are able to establish balance between self-renewal and differentiation, thereby maintain the spermatogenesis. The aim of this study was to investigate effects of different doses of Testosterone on SSCs colony formation.
The cells were isolated from testes of prepubertal lambs by two-step enzymatic digestion and then purified by differential platting. The cells were cultured for 13 days in 4 groups: Control [DMEM containing antibiotic (1%) and FBS (5%)]; and Treatment groups 1, 2 and 3 (DMEM䷫ⶢ쭞꺉 60, 120 and 240 µg/ml, respectively). The culture media was changed every 72 h. Identification of SSCs was performed by immunocytochemistry staining against PGP9.5. Finally, following the evaluation of percentage for viability rate of SSCs immediately after isolation and also the number and surface areas of the colonies on days 5, 9 and 13 after the beginning of culturing by invert microscopy, the analyses were made using statistical tests.
The percentage for viability rate of SSCs after isolation was 86.77±3.63%. The decrease in the number and surface areas of spermatogonial colonies in Treatment groups 2 and 3 were significant compared to Control and Treatment group1 (P The results of this study showed that Testosterone has no effect on colony induction of SSCs in vitro. Increasing the Testosterone dose decreases the colony number and surface area of SSCs.

Dexamethsone is one of the widely used glococorticoids that prescribe in many situations such as pregnancies that are prone to preterm delivery, so this study was conducted to investigate the effect of prenatal dexamethasone administration on the function of pituitary-gonadal axis and secretary function of testis of offspring of rats.
Material and In this study, 40 pregnant rats were divided into control group, sham group, and 3 experimental groups that receiving 0.5, 1, and 2 mg/kg doses of dexamethasone administered from the eighth day until the end of pregnancy Quaqu on day (QOD). After delivery, neonates and mothers received no treatment and after puberty, sampling from the heart and testis of male children was done, then plasma levels of testosterone,FSH, LH and the number of spermatogony, spermatocyte, spermatid, sertoli and leydig cells were determined. Results were analysed with the SPSS-20 and ANOVA and Duncan test (p≤0.5).
Results of this study revealed that, dexamethson administration leads to increase of FSH and, LH levels and decrease of testosterone levels and number of spermatogony, spermatocyte, spermatid and leydig cells.
Dexamethasone causes permanent disorders in reproductive system of male rats by inhibiting mitosis activity, increasing putative gonadotropin inhibitory hormone and impairing fetal testise development.

Infertility in men is mainly caused by as low or abnormal spermatogenesis and hormonal problems. Because of high costs and potential side effect of routine therapeutic modalities. It seems that use of herbal medicine have fewer side effects; the aim of the study is to examine the effect of palm pollen extract on serum levels of hormones and spermatozoa dynastic cells in adult male mice.

This experimental study was conducted on 40 adult male rats. They were divided into 5 groups 8, in each group that included controls (no treatment), only saline treated and three experimental groups that received aqueous extract of date palm pollen in three different dosages, doses 100, 200 and 400 mg / kg respectively. All prescriptions were done intraperitoneally for 35 days. At the end of experiment animals were phlebotomized from the heart and by separation of serum samples, amounts of testosterone, estrogen, progesterone were counted, and by separation of testicle, the numbers of spermatogonia, spermatocytes and spermatid were counted and the results were analyzed using unilateral ANOVA and Duncan methods.

The results showed that aqueous extracts of palm pollen dose-dependently caused significant increasing the levels of estrogen, progesterone and testosterone and increasing the numbers of spermatocytes and spermatid.

The presence of compounds such as cadmium, zinc, steroids, flavonoids, saponins, lipids, and by substances that inhibiting the enzyme 5- alpha redoctase could increase testosterone and sexual dynastic cells.

Preparing a suitable condition for in vitro culture of spermatogonial stem cells can help discover all of the molecular mechanisms involving in spermatogenesis and pave the way for gene manipulation. This study aimed to evaluate the effect of a poly L-lactic acid (PLLA) nanofiber on the proliferation of frozen-thawed bovine spermatogonial stem cells in vitro. The isolated spermatogonial cells from the prepubertal bull were frozen-thawed and cultured in two groups: the control group (C) including the spermatogonial cells and the treatment group (T) including the spermatogonial cells seeded onto PLLA. The cells in both groups were cultured in DMEM supplemented with 5% fetal bovine serum and 40 ng/ml glial cell line-derived neurotropic factor (GDNF) for 2 weeks. Colony assay was conducted on days 4, 7, 10 and 13 after the beginning of the culture. Expression of spermatogonial genes (PLZF, BCL6, GFRα-1, VASA, Itgα6) in the culture was determined by reverse transcription polymerase chain reaction (RT-PCR) on the last day of the experiment. The viability rates of fresh cells before and after the addition of cryoprotectant agent were 87.4±2.4 and 81.8±3.1, respectively which decreased to 65±1.7 after the thawing (P<0.01). Moreover, the results showed that the surface area of colonies derived from spermatogonia were significantly greater in the treatment group compared to the control group on day 13 (P<0.05). Finally, RT-PCR revealed the expression of all spermatogonial stem cell (SSC) markers in both groups. The PLLA nanofiber can provide a suitable microenvironment for frozen-thawed SSCs in an in vitro-culture system.

Hops (Humulus lupulus L.) has industrial and medical applications and is used in the treatment of several diseases. Infertility is a medical important issue that its treatment with chemical medicines has various side effects. Due to fewer side effects of herbal medicines, yet little research has been done on the hops. Therefore, this study aimed to evaluate the effect of hops extract on sexual hormone levels and sexual dynastic cells in Syrian adult male mice. This experimental study was performed on 40 Syrian adult male mice that were divided into 5 groups of 8: two controls groups and three experimental groups receiving various doses of hops extract (50, 100 and 150 mg/kg). Administrations were done by gavage for 35 days. At the end of the treatment period, blood samples were taken from the heart of animals and testosterone, estrogen and progesterone levels was measured. Also, after isolation of mouse testis, the number of spermatogonia, spermatocytes and spermatid were counted. Data were analyzed using ANOVA and Tukey statistical tests. The results showed that hop caused a significant increase in estrogen and testosterone levels and spermatogonia and spermatocytes cells number; but has no effect on progesterone levels. Hops extract, possibly by having phytoestrogen compounds and by stimulating LH secretion, increases estrogen and testosterone levels, and spermatogonia, spermatocytes and spermatid cells number. Therefore, further investigation on hops can utilize to help infertile men.

Spermatogonial stem cells (SSCs) are undifferentiated cells with special capabilities on the self-renewing and differentiation. Proliferation of SSCs is a primary requirement for the study of their characteristics and function in vitro. The aim of this study was to evaluate the effects of different concentrations of follicle stimulating hormone (FSH) on the colonization activity of ovine SSCs in a short-term in-vitro co-culture with sertoli cells. Both sertoli and spermatogonial cells were isolated from 2-3 months old lamb testes by two-steps enzymatic digestion. Afterward، isolated cells were cultured in four groups with different concentrations of FSH (0، 5، 10 and 15 IU/ml، respectively) for 10 days. Colony assay (number and surface area) was evaluated 4، 7 and 10 days after the beginning of the culture by light microscope. At the day 4، colony surfaces of groups 1 and 2 were significantly more than groups 3 and 4 (P < 0. 05). At the days 7 and 10، colony surfaces of groups 1 and 2 were significantly more than group 4 (P < 0. 05). The results showed that treated groups with 10 and 15 IU/ml of FSH significantly decreased colony surface of ovine SSCs in comparison with co-culture system.

Stress has different effects on the living organisms. In this study the effect of immobilization stress the HPG (hypothalamic - pituitary - gonad axis) hormones secretion and the number of spermatogonia was studied in adult male rats. 30 male rats were divided into 3 groups. The Control group: who were not exposed to any kind of stress، and two experimental 7 and 14 day groups which respectively received 2 hours of immobilized stress over 7 and 14 day stress periods، blood samples were collected directly from the heart of the animals، Hormones GnRH، LH and testosterone levels were measured by ELISA. The number of spermatogonia in the testes of adult male rats were counted. We used ANOVA and Duncan''s test results in p ≤0. 05 for statistic analyze test. The results showed that immobilization stress significantly decreased concentrations of GnRH and testosterone hormones compared with the control group. Also the 14-day immobilization stress caused a significant reduction in spermatogonia cells. The results of this study indicate that the inhibitory effect of immobilization stress on the HPG axis may also cause negative effects on the spermatogenesis process.

The increase of stress following the technological improvements appears to be an important factor that causes organs disorders like genital system. Therefore the present study was conducted to investigate the effects of chronic multiple sequential stress on rat testis 18 Wistar rats were divided randomly into two equal groups. In animals under stress، the mice were exposed to different multiple sequential stress as Forced swimming، Restraint، Water deprivation، Isolation and Food deprivation for 10 days while the animals in control group were kept in their cages without any disorders. After weighing the animals and anesthesing with xylazine-Ketamine، the testis of animals were removed and weighed. After fixation with formaldehyde (10%)، testis samples were prepared for light microscopic study. The number of spermatogonia، spermatocytes I & II، sperm & leydig cells، thickness of the seminiferous tubule determined using Image Tool software in the studied groups. Finally the data were compared statistically. The present study showed that the mean number of spermatogonia، spermatocyte I & II، sperm & leydig cells in stress group have significantly decreased compared to control group (P<0. 001). Moreover، the mean thickness of seminiferous tubule in stress group have significantly decreased compared to control group (P<0. 001) Our study showed that chronic multiple sequential stress can have negative effects by reducing the number of spermatogonia، spermatocyte I & II، sperm، leydig cells & thickness of seminiferous tubule in rat testis but more studies are needed to confirm these results.

Sesame seeds contain large amounts of antioxidants and phytoestrogens، and it has been shown that the leaf extract of this plant may have some beneficial effects on the reproductive parameters of male rats. Thus we tested the effects of the sesame oil on these parameters of reproductivity in male rats.

Fifteen mature male Wistar male rats were divided into the control and sesame oil groups. For eight weeks the control and sesame oil groups were fed the basic rat diet and basic rat diet supplemented with 5% sesame oil respectively. Following blood collection and euthanasia the epididymal sperm were counted، the morphology of testes was accessed، and leydig، sertoli، spermatogonia and spermatocyts cells were counted in histological sections of the testes. The level of testosterone and estradiol 17- β were measured.

Consumption of 5% sesame oil compared to control group، decreased blood glucose and increased the epididymal sperm count and progressive motility and the number of spermatogonia of seminiferous tubule (P<0. 05)، but had no effect on weight and testicular morphology.

This study showed that the sesame oil consumption improves some reproductive parameters، which may be related to the antioxidative and phytostrogenic properties of the sesame oil or insulin action improvement.

This study presents a simple method for isolation, expansion and purification of neonatal mouse spermatogonial stem cells. We used enzymatic digestion to isolate a cell suspension of spermatogonia and Sertoli cells from neonatal 2-day-old mice. The cells were cultured in DMEM/F12 that contained 10% serum for two weeks. Sertoli and spermatogonia cell characteristics were confirmed by examining for the presence of vimentin and PLZF proteins, respectively. To assess the rate of spermatogonia stem cell expansion, the area and number of colonies were measured during the two weeks of culture. At the end of the second week, we detected spermatogonia cell-specific expressions of the Stra8, Piwill2, DAZL, and Mvh genes. Current results indicated that isolated Sertoli and spermatogonia cells were immunopositive for specific markers. During the culture period, a significant difference was seen in the number and area of spermatogonial stem cell colonies (P<0.05) at four time points. In addition, spermatogonial specific gene expression demonstrated that these cells were undifferentiated after two weeks of culture. Our study showed that co-culture of spermatogonia and Sertoli cells from same source provides a convenient and efficient environment. This co-culture, without the addition of external growth factors and chemical manipulations, can be used for proliferation of spermatogonia stem cells.

Diazinon (DZN) and Hinosan, two of the most important organophosphate pesticides, are widely used for pest control in gardens and agriculture. These compounds adversely effect enzyme activation and reproductive organs. Therefore, in the present study, we investigated DZN and Hinosan impacts on the structure of testis tissue and sexual hormones in mice. For this study, 60 male Balb/C mice were divided into the following groups: DZN, Hinosan, control and sham. In the experimental groups, mice were injected with a single dose of DZN (0 mg/kg intraperitoneal) and Hinosan (0 mg/kg intraperitoneal), sham (corn oil) and control (no injection). Animals were sacrificed 5 days after the latest injection. Blood samples were collected and testosterone, luteinzing hormones (LH) and Follicle stimulating hormone (FSH) levels were assayed. Testis tissues sections were provided to investigate the changes of spermatogenic and Leydig cells. Testis and seminiferous diameter were assayed with micrometer and eye piece, respectively. Data were analyzed with one-way ANOVA. Significance was set at p<0.05. In the present study, no significant differences in testis and seminiferous diameter and number of blood vessels were noted. However the number of germ cells, spermatocysts, spermatids and Leydig cells on the testes of mice decreased (p<0.05). There was no significant difference between the parameters of the sham group compared to the control group. These results suggest that DZN and Hinosan can exert a decrease in sperm production. Therefore, application of DZN should be limited to a designed program.

Hinosan is an organophosphate that inhibit acetylcolinesterase activity, which could be resulted in damages of genital organs. This compound has been used extensively in the agriculture, for pest control. Therefore, in the present study we investigated the effect of Hinosan on spermatogenesis in mice. For this experimental study, the male mice were divided into three groups. In the cases group, mice were injected with Hinosan consecutive doses (20mg/kg i.p, five consecutive days per week for one month), sham (normal saline) and control (no injection). Animals were scarified 7 days after the latest Hinosan injection. Therefore, the mice testis sections were prepared and morphologic aspects of testis and spermatogenesis processes were examined. Data were analyzed using of one-way ANOVA. Significance was set at P<0.05. The Hinosan showed a significant decrease in number of germ cells, spermatocyt, spermatids, Leydig cells, blood vessels and also diameter of seminiferous on testes of the mice decreased, compared with control groups (P<0.05). This study demenstrated that Hinosan is effective on spermatogenesis and seminiferous tubule structure, also can decrease germinal cells.

Finasteride, a synthetic 4-azasteroid compound, is a competitive and specific inhibi-tor of type II 5--reductase, an intracellular enzyme that converts testosterone into dihydrotestos-terone (DHT). Finasteride is prescribed for nearly all disturbances related to DHT concentration such as benign prostatic hyperplasia, male-pattern androgenetic alopecia, hirsutism, acne and seborrhea. Since finasteride is frequently prescribed in men, the effects of different doses of finas-teride on the number of spermatogonia, Sertoli and Leydig cells have been investigated in the present study. Forty mature male Sprague-Dawley rats were divided into five groups of eight. The first group was kept as the control group and received nothing. The second or the sham group, only received distilled water orally, but the last three experimental groups respectively received 25, 50 and 100mg/kg of BW/d doses of finasteride orally for a 32-day period. Then photo-micrographs of testis tissues were studied and the results of the five groups were statistically analyzed by ANOVA, t, Tukey and Duncan tests. P<0.05 was considered significant. Administration of 50 and 100mg/kg per BW doses of finasteride significantly decreased the number of spermatogonia and 50mg/kg doses reduced the number of primary spermatocytes (p0.05). The number of Sertoli cells had no significant difference in the experimental groups in comparison with the control group but there was a significant increase in the number of Leydig cells in all of the experimental groups (p0.05). This drug did not have any significant effects on the density of different kinds of cells, and nuclear or cytoplasmic staining properties of spermato-gonia. Finasteride causes a significant decrease in the number of spermatogonia and primary spermatocytes. It also causes a significant increase in the number of Leydig cells but it does not have any significant histological effects on the testis or any effects on spermatogenesis. Therefore, it seems that short-term uses of the medication may not have harmful effects on male fertility.